Team:Berkeley Wetlab/Automation

From 2009.igem.org

Revision as of 21:21, 19 October 2009 by Jennbrophy (Talk | contribs)

Contents

Why Automate?

Our approach to characterizing cell surface display required us to make a lot of parts. Without an automated mode of assembly it would not be possible for us to make enough parts to characterize cell surface display using a combinatorial method.

Making Basic Parts

Making basic parts is the first step toward making any synthetic biology device. After designing all of the basic parts we needed for our cell surface display project, our team developed a method for doing cheap in-house gene synthesis. This method allowed us to easily make a very large number of basic parts.
(basic parts making cartoon)

oligoDesigner

We created a software called oligoDesigner to facilitate gene synthesis. OligoDesigner will take a list of goal part sequences as input and output all of the oligo nucleotides needed to synthesize the goal part. OligoDesigner will also output an IDT ([http://www.idtdna.com/Home/Home.aspx Integrated DNA Technologies]) order form that will order all of the oligo nucleotides it designed.

oligoDesigner

oligoDesigner

IDT Ordering Form

IDT Ordering Form

(IDT 96 well plate image)
BerkeleyLiquidRobot.jpg

Liquid Handling Robot

(basic parts cartoon)

The oligo nucleotides arrive from IDT in a 96 well plate format. OligoDesigner will then generate commands for a liquid handling robot to resuspend the oligo nucleotides to the correct concentrations. It will also generate commands that tell the liquid handling robot how to mix the oligos in order to get the correct PCR products.
After the liquid handling robot finishes mixing oligo nucleotides, all we need to do is add the correct buffers and enzymes and put the resultant tubes in a thermocycler for PCR.

Making Composite Parts

The next step, after making new basic parts, is to assemble those basic parts into composite part devices. There has been a lot of work being done at UC Berkeley to automate the process of composite part assembly. This year our team was able to implement many newly developed techniques to create a large number of composite parts.

Two AntiBiotic Assembly

Our team used a method of basic part assembly called Two AntiBiotic (2ab) Assembly to create basic parts.

BerkeleyAssembly.png

Clonebots


Clotho

BerkeleyAssemblytree.jpg

Success!

Using our newly developed automated assembly processes we were able to make over 800 parts (both basic and composite) in one summer! This magnitude of part making is unprecedented in iGEM competitions and enabled us to test a very large number of variations on cell surface display systems.

Success Rates

The assembly scheme we developed works just as well as assembly by hand and is much faster.

BerkeleyAssemblyScheme.jpg We made a lot of parts with a robot.